The Second Floor update gives Animal Hospital a whole new area to explore above the wards. Alongside a staff break room, the update adds a Laboratory built around a brand-new DNA experimentation system, letting players sample patients, grow cultures, and brew vials with unpredictable effects. Here is everything you need to unlock the floor, craft your first vial, and track down the secret hiding underneath it all.
Once you have completed your first ambulance call (or started with Fast Start), head to the stairs marked "UPPER FLOOR" near Aisles 6-8. Grab the DNA Sampler in the Lab, sample a checked-in patient, run the sample through the Sequencer and feed it with lettered ingredients, then finish it off in the Vial Press to collect your vial.
How to Unlock the Second Floor in Animal Hospital
You need to have gotten through your first ambulance call. Once that first emergency patient has come and gone, the stairs leading up become available for the rest of the run.
If you'd rather skip straight to testing the Lab, start your shift with the Fast Start option instead of the normal tutorial. Doing so unlocks the upper floor immediately, so you don't have to wait through an opening shift to reach it.
The stairwell itself is tucked into the hallway near Aisles 6 through 8, marked clearly with an "UPPER FLOOR" sign above the door. Walk up, interact with the handle, and climb the stairs.
What's Inside the Laboratory
The Lab is built around three machines that work together to turn a patient sample into a finished vial:
- DNA Sampler — a handheld tool you carry down to patients to collect samples
- Sequencer — grows a raw sample into a usable culture through a short minigame
- Vial Press — converts a finished culture into a working vial
Along the walls, four ingredient shelves feed the Sequencer during the culturing process. Each shelf is marked with a DNA letter and dispenses a different feed:
| Shelf Letter | Ingredient Dispensed |
|---|---|
| T | Purple mushrooms |
| A | Pink jelly |
| G | Milk |
| C | Ginger |
How to Make a DNA Vial
Turning a patient sample into a finished vial takes several trips between the wards and the Lab. Follow these steps in order:
- Grab the DNA Sampler from the counter in the Lab. Make sure it reads as ready before heading back downstairs.
- Find a checked-in patient. The sampler won't work on empty rooms, so run a normal reception cycle first and let someone actually get admitted.
- Sample the patient with the tool. Trying to skip this step and jump straight to the syringe just triggers an on-screen warning telling you to sample a patient first.
- Carry the sample back upstairs and load it into the Sequencer. This starts a short minigame — click while the marker sits inside the green zone of the DNA gauge to lock it in.
- Feed the growing culture using the lettered shelves. This is where the real depth is: certain letter sequences grow a stable culture, while others fail outright. A repeated chain like A-A-A is a known stable combination, but experimenting with your own patterns is part of the fun.
- Press the finished culture in the Vial Press and collect your vial.
- Return the DNA Sampler to the counter so it's ready for your next sample.
If the Culture Goes Unstable
An unstable chain can't be salvaged mid-process. You'll need to start the whole sequence over from a fresh sample, which means another trip down to a patient with the sampler.
What Do the Vials Actually Do?
Once you've got a finished vial, using it on a patient triggers a random effect based on the serum you brewed. Effects reported so far include:
- Shrinking a patient down to a tiny size
- Rapidly growing hair on a patient
- Knocking a patient out cold
Since a fainted patient just means more cleanup work for you, it's smart to test unknown vials during a quieter moment rather than in the middle of a packed shift.
The Secret Hatch and the Clown in the Basement
Tucked into a corner of the Lab floor is a hatch that's easy to miss. Drop through it and you'll land in a small hidden jail cell holding a clown-like NPC. Talk to him, and he'll ramble about balloons, including a strange claim about spending eighteen years "with balloons and the Laboratory."
Eventually the clown disappears, leaving behind an Animal News clipping. Picking it up completes the hidden "Clown" discovery. The clipping reveals that the Laboratory and several patient rooms were sealed off years ago with no reopening date given, alongside an unrelated report about a fatal collision involving a Nom Aly Burgers delivery truck.
Everything Else Added in the Second Floor Update
Beyond the Lab, the Second Floor patch brought a handful of smaller additions:
| Addition | What It Does |
|---|---|
| Effects board | Displays stackable status effects currently active on patients |
| New skins | Four new cosmetic skins added to the game |
| New achievements | Additional achievements tied to the Lab and Second Floor content |
| Maintenance | General bug fixes plus a leaderboard reset |
Gearing Up Faster for Harder Shifts
Time is exactly what runs short once shifts start getting chaotic. If you'd rather spend that time exploring the Lab instead of grinding out Animal Coins the slow way, the Animal Hospital shop on BLOXB sell Animal Coin bundles that go straight toward unlocking classes and other progression options.
Final Thoughts
The Second Floor update is easily the most substantial thing to happen to Animal Hospital in a while. The DNA vial system in particular stands out because it's a genuine experimentation mechanic rather than another item to simply collect, and with the newspaper hinting at sealed-off wards elsewhere in the building, this probably isn't the last new area we'll be exploring. For now, head up those stairs near Aisles 6-8, start sampling patients, and see what your first culture grows into.
Frequently Asked Questions
How do you unlock the Second Floor in Animal Hospital?
The Second Floor unlocks after your first ambulance visit. If you skip the tutorial with Fast Start, it becomes available from the very start of your shift instead.
Where are the stairs to the upper floor?
The stairwell is located in the corridor around Aisles 6 to 8, marked with an "UPPER FLOOR" sign above the door.
How do you make DNA vials in Animal Hospital?
Sample a checked-in patient with the DNA Sampler, load the sample into the Sequencer and clear the minigame, feed the growing culture with lettered ingredients until it stabilizes, then press the finished culture in the Vial Press to collect your vial.
What happens if a DNA culture becomes unstable?
An unstable culture can't be recovered. You have to start the entire process again with a fresh sample from a patient.
What do DNA vials do to patients?
Using a finished vial on a patient triggers a random effect, such as shrinking them down, making hair grow rapidly, or knocking them out cold.
Where is the secret clown room in Animal Hospital?
It's accessed through a hidden hatch in one corner of the Laboratory floor, which drops down into a small jail cell holding a clown-like NPC.
Can you buy Animal Coins to progress faster?
Yes. Sites such as the Animal Hospital shop on BLOXB sell Animal Coin bundles that can be spent on classes and other progression options, though it isn't required to enjoy the Second Floor update.